Related Experiment Video
Updated: Jun 19, 2026
10:25
Using the E1A Minigene Tool to Study mRNA Splicing Changes
Published on: April 22, 2021
Exon sequences at the splice junctions affect splicing fidelity and alternative splicing
Luciana B Crotti1, David S Horowitz
1Department of Biochemistry and Molecular Biology, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Rd., Bethesda, MD 20814, USA.
Summary
Exon sequences at splice junctions significantly influence 3' splice site selection during pre-mRNA splicing. This finding, observed in yeast, impacts splicing fidelity and alternative splicing.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Accurate pre-mRNA splicing is crucial for eukaryotic gene expression.
- Splice sites are recognized by the splicing machinery based on specific sequences.
- The role of exon sequences in splice site selection is not fully understood.
Purpose of the Study:
- To investigate the role of exon sequences in 3' splice site selection.
- To determine the impact of exon sequences on splicing fidelity and alternative splicing.
- To explore the relationship between splicing signals and the genetic code.
Main Methods:
- In vitro splicing assays using yeast.
- Kinetic analysis of splicing reactions.
- Point mutations in exon sequences of yeast genes.
Main Results:
- Exon sequences at splice junctions play a significant role in 3' splice site selection during the second step of splicing.
- The effect of exon sequences was enhanced in a Prp18 mutant (Prp18DeltaCR).
- Point mutations in exon bases altered splice site selection, affecting splicing fidelity and alternative splicing in yeast.
Conclusions:
- Exon sequences are critical for accurate 3' splice site selection.
- Splice site selection occurs competitively during the second step of splicing.
- The evolution of splicing signals and the genetic code may be interconnected.
Related Concept Videos
02:18Alternative RNA Splicing
Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
02:18Alternative RNA Splicing
Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
01:32RNA Splicing
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
01:32RNA Splicing
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
02:41Chromatin Structure Regulates pre-mRNA Processing
In eukaryotic cells, nascent mRNA transcripts need to undergo many post-transcriptional modifications to reach the cell cytoplasm and translate into functional proteins. For a long time, transcription and pre-mRNA processing were considered two independent events that occur sequentially in the cell. However, it has now been well established that transcription and pre-mRNA processing are two simultaneous processes that are precisely regulated inside the cell.
The chromatin structure, especially...
The chromatin structure, especially...
01:32Pre-mRNA Processing: RNA Splicing
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...